PUBLICATION

Infection of zebrafish larvae with human norovirus and evaluation of the in vivo efficacy of small-molecule inhibitors

Authors
Van Dycke, J., Cuvry, A., Knickmann, J., Ny, A., Rakers, S., Taube, S., de Witte, P., Neyts, J., Rocha-Pereira, J.
ID
ZDB-PUB-210411-2
Date
2021
Source
Nature Protocols   16(4): 1830-1849 (Review)
Registered Authors
de Witte, Peter
Keywords
none
MeSH Terms
  • Animals
  • Caliciviridae Infections/virology*
  • Embryo, Nonmammalian/virology
  • Humans
  • Immunity, Innate
  • Larva/drug effects
  • Larva/immunology
  • Norovirus/genetics
  • Norovirus/physiology*
  • Reproducibility of Results
  • Small Molecule Libraries/pharmacology*
  • Virus Replication
  • Zebrafish/embryology
  • Zebrafish/virology*
PubMed
33837302 Full text @ Nat. Protoc.
Abstract
We have recently established that human norovirus (HuNoV) replicates efficiently in zebrafish larvae after inoculation of a clinical sample into the yolk, providing a simple and robust in vivo system in which to study HuNoV. In this Protocol Extension, we present a detailed description of virus inoculation by microinjection, subsequent daily monitoring and harvesting of larvae, followed by viral RNA quantification. This protocol can be used to study viral replication of genogroup (G)I and GII HuNoVs in vivo within 3-4 d. Additionally, we describe how to evaluate the in vivo antiviral effect and toxicity of small molecules using HuNoV-infected zebrafish larvae, in multi-well plates and without the need for specific formulations. This constitutes a great advantage for drug discovery efforts, as no specific antivirals or vaccines currently exist to treat or prevent norovirus gastroenteritis.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping